Objective To explore the protective effect of olfactory ensheathing cell(OEC)Schwann cell(SC)-extracellular matrix(ECM)-poly(DL-lactide-co-glycolide acid)(PLGA)bridging complex on peripheral target organ and spinal cord neurons after rat sciatic nerve defect.Methods A 15 mm right sciatic nerve defect model was established in SD rats and repaired with OEC-SC-ECM-PLGA bridging complex that contained OEC,SC,ECM and self-made PLGA conduit.At the same time,the study set OEC-ECM-PLGA group,SC-ECM-PLGA group,ECM-PLGA group,PLGA group and nerve autograft control group.At 1,3,6 and 9 weeks after surgery,the gastrocnemius muscle water weight test and motor end-plate test were performed.At the 9th week after surgery,CM-DiI and horseradish peroxidase(HRP)retrograde tracing were performed.Results The gastrocnemius muscle water weight and number of motor end-plate were decreased in all groups after surgery but gradually increased after three weeks except for ECM-PLGA group and PLGA group.At the 9th week,OEC-SC-ECM-PLGA group showed no statistical differences with nerve autograft group in aspects of gastrocnemius muscle water weight,number of motor end-plate and length of motor end-plate major axis(P ＞ 0.05).At the 9th week,CM-DiI and HRP retrograde tracing found that the number of positive neurons in spinal cord in OEC-SC-ECM-PLGA group showed no statistical differences compared with nerve autograft group(P ＞0.05).Conclusions OEC-SC-ECM-PLGA bridging complex can partially protect peripheral target organ and spinal cord neurons after rat sciatic nerve defect.

Objective To determine the role and mechanism of peroxisome proliferator activated receptors (PPAR) α in a mouse model of D-galactosamine/lipopolysaccharide (D-GalN/LPS)-induced acute liver failure(ALF).Methods Firstly,C57BL/6 mice were randomly divided into control group(n =8),ALF 2h group(n =8),ALF 4h group (n =8),ALF 6h group (n =8).Secondly C57BL/6 mice were randomly divided into control group(n =8),ALF group(n =8),WY14643 group(n =8).To induce ALF,the mice were injected intraperitoneally with D-GalN (700 mg/kg) and LPS (10 μg/kg).WY14643 (6 mg/kg),the selective agonist of PPAR α,was administered via tail vein two hours prior to D-GalN/LPS exposure.Two,four,and six hours after D-GalN/LPS treatment in the first study,mice were anesthetized and blood was collected,6h after D-GalN/LPS treatment in the second study,blood was collected.The liver tissue was harvested for histology and mRNA extraction.Serum levels of ALT and AST were measured to evaluate the hepatic damage.Inflammatory cytokines (TNFα,IL-1β,IL-6) and chemokines (CXCL-1,CXCL-10) were detected by real-time quantitative PCR.Differential protein expression of p-NF-κBp65,p-JNK,p-ERK,p-p38 in inflammatory pathways was detected by Western blotting.Significance of inter-group differences was assessed by one-way ANOVA,and pairwise comparison was performed by the least significant difference test.Results The gene and protein expression of PPAR α were gradually reduced during the development of ALF.Compared with the model group,the liver architecture was better preserved almost with normal morphology in WY14643-treated mice.Serum ALT and AST levels in WY14643-treated group were significantly lower [ALT:(555 ±62)U/L vs (2 898 ±822) U/L,P ＜0.05; AST:(791 ±58) U/L vs (3 013 ±997)U/L,P ＜ 0.05].The expression of proinflammatory cytokines and chemokines was significantly suppressed during the activation of PPAR α.In the second study,the levels of gene expression of proinflammatory cytokines and chemokines were detected in control group,ALF group and WY14643 group respectively as followings:TNFα (0.161 ± 0.085,7.996 ± 1.068,3.346 ± 0.94,P ＜ 0.05),IL-1β(0.041 ±0.002,3.657 ±0.904,0.176±0.089,P＜0.01),IL-6 (0.018 ±0.008,1.762 ±0.589,0.163±0.0487,P ＜0.05),CXCL-1 (0.063 ±0.008,7.881 ±0.966,2.737 ±0.864,P ＜0.01),CXCL-10 (0.054 ±0.005,5.671 ±0.948,2.578 ±0.804,P ＜0.05).Conclusion Our findings first demonstrate that PPARα protects liver from injury in an ALF mouse model by suppressing inflammatory response,indicating PPARα as a potential new therapeutic target for ALF.

Objective To detect γ-secretase gene mutations in a large Chinese pedigree with acne inversa (AI).Methods Clinical evaluation was carried out in a large pedigree with AI through field investigation.Peripheral blood samples were obtained from 17 family members (11 affected and 6 unaffected) and 100 unrelated healthy human controls.DNA was extracted from the blood samples,and PCR was performed to amplify all the coding regions of PSEN 1,PSENEN and NCSTN genes followed by DNA sequencing analysis.Results There were 67 members over 5 generations in this family,of whom,25 (13 males and 12 females) were affected by AI.AI was inherited in an autosomal dominant manner in this family.Skin lesions were mainly distributed on the neck,back,chest and buttocks,and occasionally in subaxillary regions.DNA sequencing revealed a novel missense mutation,c.1258C＞ T (p.Q420XP),in the exon 11 of the NCSTN gene in 11 affected family members,which leads to a substitution of glutamine by a premature termination codon at amino acid 420 (p.Q420X).The mutation was undetected in either the unaffected members or the unrelated healthy controls,and had not been registered in the single nucleotide polymorphism (SNP) database in National Center for Biotechnology Information.Conclusions There is a novel heterozygous missense mutation,c.1258C ＞ T in the exon 11 of the NCSTN gene,which may be the molecular basis of pathogenesis of AI in this family.

Objective:To investigate the efficacy and safety of different doses of tirofiban combined with coronary artery intervention in treatment of Non ST-segment elevation acute coronary syndromes (NST-ASC).Methods:110 cases with NST-ACS from October 2014 to January 2016 in our hospital were chosen and divided into the all dose group and half dose group.The TIMI blood grade before and after treatment,cardiac function before and after treatment for 30 d,major adverse cardiac events and bleeding events,hospitalization expenses and hospitalization days were recorded and compared between two groups.Results:Compared with before treatment,the TIMI 2 grade and 3 grade were obvious decreased,and the left ventricular end diastolic volume (LVEDV),left ventricular end systolic volume (LVESV) after treatment for 30 d were all obvious decreased,while the left ventricular ejection fraction (LVEF) were significant increased,P＜0.05.And the TIMI blood flow grading,LVEDV,LVESV and LVEF before and after treatment in two groups had no significant difference (P＞0.05),and the major adverse cardiac events and hospitalization days of two groups had no significant difference,P＞0.05.While the bleeding events and hospitalization expense of all dose group was obvious higher than those of half dose group,P＜0.05.Conclusions:The half dose group of tirofiban combined with coronary artery intervention in treatment NST-ACS has obvious efficacy,it can decrease the bleeding events and hospitalization expense.

Objective To construct a recombinant lentiviral expression vector for RNA interference (RNAi) of human Snail gene and to study its effects on the proliferation and invasion of nasopharyngeal carcinoma cell line 5-8F. Methods The effective sequence of short hairpin RNAs (shRNA) targeting Snail gene was designed and cloned into the linear pLVTHM vector after enzyme digestion. After confirmation by DNA sequencing, 5-8F cells were infected with the viral supernatants. The cells with stable Snail gene knock-down were separated by fluorescence activated cell sorter (FASC). The expression of Snail mRNA was detected by real time RT-PCR. MTT and cell invasion assay were used to detect the proliferation and invasion of 5-8F cells after plVTHM-siSnail transfection. Results The lentivirus vector plVTHM-siSnail was constructed successfully. The separated 5-8F-plVTHM-siSnail exhibited significant knock-down of Snail mRNA expression. Slower proliferation and decreased cells to permeate through the Matrigel were found after plVTHM-siSnail transfection (P

Extensive functions can be provided to hospital management,medical treatment and teaching when massive data are reasonably processed and analyzed. Knowledge mined from sequence of doctor's advice for medicine can not only evaluate treatment quality but also provide necessary basis for establishing a safety and effective medicine treatment plan. This paper analyzes some common information of doctor's advice for medicine,mines the doctor's advice for medicine by using sequence pattern mining method,and explains the mining results.

Objective To investigate the prevalence and features of thyroid nodules in middle and aged patients with type 2 diabetes mellitus(T2DM). Methods High-resolution ultrasonography was used to detect thyroid nodules in 132 cases middle and aged patients with type 2 diabetes and 89 patients without diabetes.The nodule features and its relationships with related indicators in diabetic patients were analyzed. Results The prevalence of thyroid nodules in middle and aged patients with type 2 diabetes was higher than that without diabetes (67.4％ vs. 53.9％,P＜0.05),and most occurred in 50 to 59 age group (66.7％ vs. 42.9％) without dependence on changes in thyroid functions and volumes.In diabetes group,the prevalence of thyroid nodules were 59.5％ in male and 81.3％ in female (P＜0.05),no obvious difference was observed in the size and number of thyroid nodules between male and female,multiple nodules and micronodule (＜ 1.0 cm) had the higher incidences in both sexes.The prevalence of thyroid nodules was increased with aging,but not with diabetes duration and glycosylated hemoglobin (HbAlc) level (x2 =0.797,P=0.372; x2 =1.078,P =0.229). Conclusions It is common that thyroid nodules combined with diabetes in middle and aged patients,thyroid ultrasound screening and regular following-up of patients aged ≥50 years have important clinical significance.

Objective To discuss the combined value of gray-scale ultrasound ( GSUS) and contrast-enhanced ultrasound (CEUS) in the diagnosis of benign and malignant thyroid lesions and to explore the optimal diagnostic point of scoring method .Methods Ultrasound images of 178 thyroid lesions confirmed by pathology were synthetically reviewed by scoring 5 GSUS indicators including shape , orientation, interior echogenicity, halo sign, microcalcification and 6 CEUS indicators including relative arrival time of microbubbles in the periphery and interior, peak periphery and interior echogenicity, peripheral ring-enhancement, and homogeneity of enhancement .One positive indicator scored one point .The optimal diagnostic points and their clinical value were explored according to ROC curves .Results Scores of GSUS, CEUS and the combination of GSUS and CEUS were significantly different (Z =10.188,9.843,10.705,all P <0.001). Areas under ROC curves of GSUS, CEUS and the combination of GSUS and CEUS were 0.936, 0.919 and 0.964, respectively.Three or more positive GSUS indicators of five in a thyroid lesion predicted that the thyroid lesion was malignant , with the sensitivity of 79.6% and the specificity of 91.2%.Two or more positive CEUS indicators of six in a thyroid lesion predicted that the thyroid lesion was malignant , with the sensitivity of 91.8% and the specificity of 81.2%.Five or more positive GSUS and CEUS indicators of eleven in a thyroid lesion predicted that the thyroid lesion was malignant , with the sensitivity of 93.6%and the specificity of 92.3%.The areas under the ROC curve of GSUS and CEUS were 0.936 and 0.919.The area under the ROC curve of the combination of GSUS and CEUS was 0.964, larger than the areas under the GSUS ROC curve and the CEUS ROC curve.Conclusion Ultrasound is valuable in the differential diagnosis of benign and malignant thyroid lesions , and the combination of GSUS and CEUS is the most valuable with 5 points as the optimal diagnostic scoring method .

Objective To study the enhancing effect of ultrasound plus microbubble on small interference RNA(siRNA)transfection in vitro and in vivo.Methods Human vascular epithelial growth factor(VEGF)siRNA with 2'deoxy modification(VEGF 2'-eoxy siRNA,VdsR)was used.The human CNE cells(fromnasopharyngeal carcinoma)line was used for in vitro cell-based experiments and in vivo mouse xenograft model.Two different microbubble agents.BR14 and Levovist.were used together with the RNA transfection reagent RNA-mate.ELISA and RT-PCR assays were used to assess VEGF gene expression.Immunohistochemical staining (IHC)was performed to assess CD31 expression in xenograft tumors.Results VdsR transfection in CNE cells abolished VEGF expression as determined by ELISA experiments.In the first mouse xenograft experiment,ultrasound exposure dramatically enhanced VdsR-mediated tumor inhibition.In the second mouse xenograft experiment,when VdsR was mixed with the microbubble reagents and then injected into xenografts,ultrasoundexposure significantly reduced tumor growth in BR14-mixed VdsR group but not in the Levovist-mixed VdsR group compared to the control.RT-PCR experiments demonstrated that VEGF expression in ultrasound-exposed tumors was significantly lower than that in the control.Meanwhile,VEGF expression in the tumor tissue treated by BRl4-mixed VdsR declined as compared with the controls.Tumor vascular density as measured by CD31 immunostaining was significantly decreased in ultrasound-exposed tumors compared to the control.Conclusions Ultrasound exposure and/or microbubble can significantly enhance delivery and the efficiency of VdsR-mediated anti-tumor effects,and should be a location-specific enhancement approach for siRNA-based anti-cancer therapy.

Two fragments G1 and G2 of the glycoprotein G gene of bovine respiratory syncytial virus(BRSV) were selected for expression in Escherichia coli based on the analysis of glycoprotein G by DNA Star software.Then the two fragments of glycoprotein G were amplified by PCR with synthesized G gene of BRSV as the template.The amplified fragments G1 and G2 are 570bp and 308bp in length,respectively.The PCR products were cloned into pET30a vector and expressed in soluble form in E.coli after induction of cultured E.coli with IPTG.Both of the recombinant proteins G1 and G2 were purified by immobilized Ni ion affinity chromatography under native conditions.Then the purified proteins were analysed by Western blotting.The results showed that the purified recombinant protein G1 retained good antigenicity and specificity.But the purified recombinant protein G2 didn't possess biological activity.Antibodies against BRSV were detected in suspected bovine serum samples in China by using indirect ELISA and Western blotting with the purified recombinant protein G1.The purified recombinant protein G1 might be used as antigen for establishing serological methods for diagnosis of BRSV infection.And the purified recombinant protein G1 might also be used for preparing polyclonal and monoclonal antibodies for research on biological functions of glycoprotein G of BRSV.